The difficulty of censusing marine animal populations hampers effective ocean management. Analyzing water for DNA traces shed by organisms may aid assessment. Here we tested aquatic environmental DNA ...(eDNA) as an indicator of fish presence in the lower Hudson River estuary. A checklist of local marine fish and their relative abundance was prepared by compiling 12 traditional surveys conducted between 1988-2015. To improve eDNA identification success, 31 specimens representing 18 marine fish species were sequenced for two mitochondrial gene regions, boosting coverage of the 12S eDNA target sequence to 80% of local taxa. We collected 76 one-liter shoreline surface water samples at two contrasting estuary locations over six months beginning in January 2016. eDNA was amplified with vertebrate-specific 12S primers. Bioinformatic analysis of amplified DNA, using a reference library of GenBank and our newly generated 12S sequences, detected most (81%) locally abundant or common species and relatively few (23%) uncommon taxa, and corresponded to seasonal presence and habitat preference as determined by traditional surveys. Approximately 2% of fish reads were commonly consumed species that are rare or absent in local waters, consistent with wastewater input. Freshwater species were rarely detected despite Hudson River inflow. These results support further exploration and suggest eDNA will facilitate fine-scale geographic and temporal mapping of marine fish populations at relatively low cost.
Stress granules (SGs) form during cellular stress and are implicated in neurodegenerative diseases such as amyotrophic lateral sclerosis and frontotemporal dementia (ALS/FTD). To yield insights into ...the role of SGs in pathophysiology, we performed a high-content screen to identify small molecules that alter SG properties in proliferative cells and human iPSC-derived motor neurons (iPS-MNs). One major class of active molecules contained extended planar aromatic moieties, suggesting a potential to intercalate in nucleic acids. Accordingly, we show that several hit compounds can prevent the RNA-dependent recruitment of the ALS-associated RNA-binding proteins (RBPs) TDP-43, FUS, and HNRNPA2B1 into SGs. We further demonstrate that transient SG formation contributes to persistent accumulation of TDP-43 into cytoplasmic puncta and that our hit compounds can reduce this accumulation in iPS-MNs from ALS patients. We propose that compounds with planar moieties represent a promising starting point to develop small-molecule therapeutics for treating ALS/FTD.
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•∼100 small-molecule compounds modulate SGs in HEK293xT cells, NPCs, and iPS-MNs•ALS-associated RBPs accumulate in SGs during prolonged stress•Molecules with planar moieties disrupt accumulation of ALS-associated RBPs in SGs•Compounds reduce TDP-43 accumulation in cytoplasmic puncta in ALS mutant iPS-MNs
Using high-content screening, we identified a class of planar small molecules that can (1) modulate the dynamics of neurodegeneration-linked stress granules (SGs), (2) reduce SG association of ALS-linked RNA-binding proteins, and (3) prevent accumulation of TDP-43 within persistent cytoplasmic puncta.
Stress granules (SGs) are transient ribonucleoprotein (RNP) aggregates that form during cellular stress and are increasingly implicated in human neurodegeneration. To study the proteome and ...compositional diversity of SGs in different cell types and in the context of neurodegeneration-linked mutations, we used ascorbate peroxidase (APEX) proximity labeling, mass spectrometry, and immunofluorescence to identify ∼150 previously unknown human SG components. A highly integrated, pre-existing SG protein interaction network in unstressed cells facilitates rapid coalescence into larger SGs. Approximately 20% of SG diversity is stress or cell-type dependent, with neuronal SGs displaying a particularly complex repertoire of proteins enriched in chaperones and autophagy factors. Strengthening the link between SGs and neurodegeneration, we demonstrate aberrant dynamics, composition, and subcellular distribution of SGs in cells from amyotrophic lateral sclerosis (ALS) patients. Using three Drosophila ALS/FTD models, we identify SG-associated modifiers of neurotoxicity in vivo. Altogether, our results highlight SG proteins as central to understanding and ultimately targeting neurodegeneration.
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•APEX proximity labeling reveals ∼150 unknown SG proteins in a dense protein network•SG composition varies by stress and cell type, especially in neuronal cells•ALS motor neurons contain SGs with distinct content and subcellular distribution•SG proteins modify ALS-mutant-mediated toxicity in fly models of neurodegeneration
Interactions between stress granule proteins exist ahead of a stress response and candidate SG proteins modify disease phenotypes in vivo.
BACKGROUNDThe rod and frame test (RFT), a measure of field dependence-independence, recently has reemerged as a measure of research interest and potential diagnostic value in neuropsychology. In the ...standard RFT, the subject experiences offsetting visual cues from a frame surrounding an embedded rod, while the subject's postural/vestibular cues provide the sense of verticality as the subject attempts to set the rod to vertical. The paper shows that RFTs not adhering to RFT parameters can reduce the test's visual framework impact experienced by the subject. Comparisons of neuropsychological studies will highlight that correct adherence to RFT testing conditions can strengthen RFT effects.METHODThis review presents the parameters that have been studied which impact on subject performance on the RFT. It identifies how computer administered RFTs have been applied to enhance the study of the RFT parameters and make the RFT more accessible to the study of different diagnostic groups. The article also critiques studies by identifying how the RFT's parameters, study's design and statistical analysis may have diminished identifying the full effects of the RFT experience.RESULTSParameters impacting judgments of verticality of the rod can include: perceived size of rod and frame, the gap between the ends of the rod and surrounding frame, presentation of the rod within an encompassing 3D visual framework that visually blocks out the surrounding environment, a dark room, instructions stressing egocentric vs allocentric strategies, double frame surrounding the rod to assess global perception effects, etc. Details are presented how gap size likely affected results in neuropsychology studies. Potentially, these and other experiments may be studied using computer administered RFTs.CONCLUSIONSBased on the descriptions of computer administered RFTs, this article suggested that incorporating these technologies can provide better understanding underlying the RFT, and in turn, understanding neuropsychology processes.
Informal recycling is a new and expanding low cost recycling practice in managing Waste Electrical and Electronic Equipment (WEEE or e-waste). It occurs in many developing countries, including China, ...where current gaps in environmental management, high demand for second-hand electronic appliances and the norm of selling e-waste to individual collectors encourage the growth of a strong informal recycling sector. This paper gathers information on informal e-waste management, takes a look at its particular manifestations in China and identifies some of the main difficulties of the current Chinese approach. Informal e-waste recycling is not only associated with serious environmental and health impacts, but also the supply deficiency of formal recyclers and the safety problems of remanufactured electronic products. Experiences already show that simply prohibiting or competing with the informal collectors and informal recyclers is not an effective solution. New formal e-waste recycling systems should take existing informal sectors into account, and more policies need to be made to improve recycling rates, working conditions and the efficiency of involved informal players. A key issue for China’s e-waste management is how to set up incentives for informal recyclers so as to reduce improper recycling activities and to divert more e-waste flow into the formal recycling sector.
Abstract
Environmental DNA (eDNA) technology potentially improves the monitoring of marine fish populations. Realizing this promise awaits better understanding of how eDNA relates to fish presence ...and abundance. Here, we evaluate performance by comparing bottom trawl catches to eDNA from concurrent water samples. In conjunction with New Jersey Ocean Trawl Survey, 1-l water samples were collected at surface and depth prior to tows at about one-fourth of Survey sites in January, June, August, and November 2019. eDNA fish diversity from 1 l was same as or higher than trawl fish diversity from 66 M litres swept by one tow. Most (70–87%) species detected by trawl in a given month were also detected by eDNA, and vice versa, including nearly all (92–100%) abundant species. Trawl and eDNA peak seasonal abundance agreed for ∼70% of fish species. In log-scale comparisons by month, eDNA species reads correlated with species biomass, and more strongly with an allometric index calculated from biomass. In this 1-year study, eDNA reporting largely concorded with monthly trawl estimates of marine fish species richness, composition, seasonality, and relative abundance. Piggybacking eDNA onto an existing survey provided a relatively low-cost approach to better understand eDNA for marine fish stock assessment.
RNA-programmed genome editing using CRISPR/Cas9 from Streptococcus pyogenes has enabled rapid and accessible alteration of specific genomic loci in many organisms. A flexible means to target RNA ...would allow alteration and imaging of endogenous RNA transcripts analogous to CRISPR/Cas-based genomic tools, but most RNA targeting methods rely on incorporation of exogenous tags. Here, we demonstrate that nuclease-inactive S. pyogenes CRISPR/Cas9 can bind RNA in a nucleic-acid-programmed manner and allow endogenous RNA tracking in living cells. We show that nuclear-localized RNA-targeting Cas9 (RCas9) is exported to the cytoplasm only in the presence of sgRNAs targeting mRNA and observe accumulation of ACTB, CCNA2, and TFRC mRNAs in RNA granules that correlate with fluorescence in situ hybridization. We also demonstrate time-resolved measurements of ACTB mRNA trafficking to stress granules. Our results establish RCas9 as a means to track RNA in living cells in a programmable manner without genetically encoded tags.
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•RNA-targeting Cas9 (RCas9) enabled recognition of endogenous, unmodified mRNAs•RCas9 did not influence mRNA abundance or amount of translated protein•Subcellular distribution of RCas9 was highly correlated with RNA-FISH•RCas9 revealed trafficking of mRNAs to stress granules in live cells
RNA-targeting Cas9 enables tracking of endogenous, untagged mRNA, establishing CRISPR/Cas9 as a programmable system to recognize RNA in live cells.
The high susceptibility of humans to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, the cause of coronavirus disease 2019 (COVID-19), reflects the novelty of the virus and ...limited preexisting B cell immunity. IgG against the SARS-CoV-2 spike (S) protein, which carries the novel receptor binding domain (RBD), is absent or at low levels in unexposed individuals. To better understand the B cell response to SARS-CoV-2 infection, we asked whether virus-reactive memory B cells (MBCs) were present in unexposed subjects and whether MBC generation accompanied virus-specific IgG production in infected subjects. We analyzed sera and peripheral blood mononuclear cells (PBMCs) from non-SARS-CoV-2-exposed healthy donors and COVID-19 convalescent subjects. Serum IgG levels specific for SARS-CoV-2 proteins (S, including the RBD and S2 subunit, and nucleocapsid N) and non-SARS-CoV-2 proteins were related to measurements of circulating IgG MBC levels. Anti-RBD IgG was absent in unexposed subjects. Most unexposed subjects had anti-S2 IgG, and a minority had anti-N IgG, but IgG MBCs with these specificities were not detected, perhaps reflecting low frequencies. Convalescent subjects had high levels of IgG against the RBD, S2, and N, together with large populations of RBD- and S2-reactive IgG MBCs. Notably, IgG titers against the S protein of the human coronavirus OC43 were higher in convalescent subjects than in unexposed subjects and correlated strongly with anti-S2 titers. Our findings indicate cross-reactive B cell responses against the S2 subunit that might enhance broad coronavirus protection. Importantly, our demonstration of MBC induction by SARS-CoV-2 infection suggests that a durable form of B cell immunity is maintained even if circulating antibody levels wane.
The recent rapid worldwide spread of SARS-CoV-2 has established a pandemic of potentially serious disease in the highly susceptible human population. Key issues are whether humans have preexisting immune memory that provides some protection against SARS-CoV-2 and whether SARS-CoV-2 infection generates lasting immune protection against reinfection. Our analysis focused on pre- and postinfection IgG and IgG memory B cells (MBCs) reactive to SARS-CoV-2 proteins. Most importantly, we demonstrate that infection generates both IgG and IgG MBCs against the novel receptor binding domain and the conserved S2 subunit of the SARS-CoV-2 spike protein. Thus, even if antibody levels wane, long-lived MBCs remain to mediate rapid antibody production. Our study results also suggest that SARS-CoV-2 infection strengthens preexisting broad coronavirus protection through S2-reactive antibody and MBC formation.
Immersive virtual reality (IVR) is a form of distraction therapy that has shown potential as an analgesia and sedation sparing agent. This study assessed the effect of IVR on the self-administered ...sedation requirements of patients undergoing joint replacement surgery under regional anesthesia in a single center.
This study was a single-center, randomized control trial at St Vincent's Hospital in Melbourne, Australia. Fifty patients undergoing elective total knee and total hip arthroplasty were randomized to IVR and Propofol patient-controlled sedation (PCS) or propofol PCS alone. The primary outcome measure was intra-operative propofol use. Secondary outcomes included pattern of propofol use over time, use of adjunct analgesia, unmet propofol demand, and patient satisfaction survey scores. Of 50 total patients, 25 received IVR in conjunction with PCS, and 25 received PCS alone. All patients received adjunct analgesia from the treating Anesthesiologist. Median propofol use/hour over the entire procedure in the control group was 40 (11.1, 93.9) mg/hour compared with 45 (0, 94.7) mg/hour in the IVR group (p = 0.90). There were no differences in patterns of propofol use over the course of each procedure. Adjusting for various baseline characteristics did not change the results. Postoperative satisfaction scores were equivalent in both groups. The VR intervention was well tolerated by all patients, with no report of major side effects. Key limitations were relatively small sample size, the non-blinded nature of the study, and use of adjunct analgesia.
In patients receiving joint replacement surgery under regional anesthesia with PCS, IVR was well tolerated but did not decrease the overall sedation requirement.
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